Antioxidant Activity and Flavonoids Identification by LC-MS/MS Analysis in Leaf Extract from Trattinnickia rhoifolia Willd

The objectives of this work were to evaluate the antioxidant and hepatoprotective activities in the oxidative stress model induced by paracetamol in male Swiss mice, to evaluate the hypoglycemic and hypolipidemic action and to identify flavonoids in the leaves ethanolic extract from Trattinnickia rhoifolia. By the LC-MS / MS method, eight flavonoids were identified in the hydromethanolic (HM) and ethyl acetate (EA) fractions, except for Amentoflavone, the flavonoids Apigenin, Canferol, Luteolin, Quercetin, Quercetin-3-β-d-glucoside, Rutin and Taxifoline were identified for the first time in this species. These fractions were evaluated for antioxidant capacity (DPPH test) and their protective effect in vivo through the analysis of superoxide dismutase, catalase, glutathione-s-transferase, non-protein thiols, ascorbic acid, TBARS and carbonylated proteins. The data showed that EA has antioxidant capacity and superior oxidative stress repair in chemical and biological analyzes, besides hypoglycemic and hypolipidemic action. Thus, the present work contributes significantly to the literature, since it is the first study that identifies the chemical constituents and pharmacological properties of leaves extract from T. rhoifolia.Os objetivos deste trabalho foram avaliar as atividades antioxidante e hepatoprotetora no modelo de estresse oxidativo induzido por paracetamol em camundongos Swiss machos,avaliar a ação hipoglicemiante e hipolipidêmica e identificar flavonoides no extrato etanólico de folhas da Trattinnickia rhoifolia. Pelo método de LC-MS/MS foram identificados oito flavonoides nas frações hidrometanólica (HM) e acetato de etila (EA), com exceção da Amentoflavona, os flavonoides Apigenina, Canferol, Luteolina, Quercetina, Quercetina-3-β-d-glicosídeo, Rutina e Taxifolina, foram identificados pela primeira vez na espécie. Estas frações foram avaliadas quanto à capacidade antioxidante (teste de DPPH) e seu efeito protetor in vivo através das análises de superóxido dismutase, catalase, glutationa-s-transferase, tiois não-proteicos, ácido ascórbico, TBARS e proteínas carboniladas. Os dados mostraram que EA possui capacidade antioxidante e reparo do estresse oxidativo superior nas análises químicas e biológicas, além de ação hipoglicemiante e hipolipidêmica. Assim, o presente trabalho contribui significativamente para a literatura, uma vez que é o primeiro estudo que identifica os constituintes químicos e propriedades farmacológicas do extrato de folhas de T. rhoifolia

Chemoprotective Effect of Leaf Extracts of Cecropia distachya Huber (Urticaceae) in Mice Submitted to Oxidative Stress Induced by Cyclophosphamide

The objective of this work was to evaluate the chemoprotective effects of the embaúba Cecropia distachya Huber (two methanolic fractions: F1 and F2) on oxidative stress induced by cyclophosphamide (75 mg kg-1) in mice, as well as phytochemical analyzes by LC-MS/MS. In the experimental model, the fractions increased the hepatic and cardiac catalase (CAT), reduced glutathione (GSH) of the kidney and the heart. F1 increased platelet levels (PLT), hemogram (RBC), hematocrit (HCT) and hemoglobin (Hb) and F1 reduced hepatic lipid peroxidation (TBARS) and aspartate aminotransferase (AST) activity increased. F1 was attributed to possible hypoglycemic activity and possible immunosuppressive effects and F2 presented antimutagenic activity. In some cases, the fractions were also hepatotoxic. These results demonstrated that the fractions stimulate the antioxidant defenses, being hypoglycemic (F1) and antimutagenic (F2), and, the harmful effects attributed to the fractions may be the association of compounds that were not elucidated in this work. &nbsp

Maize protein bodies proteome by LC-MS/MS and LC/LC-MS/MS

O valor nutricional do milho é limitado devido ao seu alto conteúdo de proteínas de reserva (zeínas), que são deficientes em aminoácidos essenciais como lisina e triptofano. Por esse motivo, a Embrapa (Empresa Brasileira de Pesquisa Agropecuária) desenvolveu a variedade de milho BR473, que tem um excelente valor energético e um maior conteúdo protéico quando comparado ao milho comum. Tal variedade possui 0,9 e 4,0 g/kg de grão de triptofano e lisina, respectivamente, contra 0,6 e 2,6 do milho comum. O alto conteúdo de lisina em algumas variedades de milho como a opaco-2 (o2), por exemplo, é relacionado à presença de proteínas não-zeínas. Para identificar essas proteínas, o que poderia explicar o alto conteúdo protéico da variedade BR473, foi analisado o proteoma dos corpos protéicos dos grãos desse milho. Para tal, foram usadas e avaliadas as cromatografias uni e bidimensional, acopladas a espectrometria de massas (LC-MS/MS e LC/LC-MS/MS), uma vez que tais técnicas se tornaram ferramentas poderosas para seqüenciamento e identificação de proteínas. Neste trabalho, foram identificadas seis proteínas por LC-MS/MS e dezesseis proteínas por LC/LC-MS/MS.The nutritional value of maize seed is limited due to its high content of zeins (storage proteins), which are deficient in essential amino acids as lysine and tryptophan. Because of this, Embrapa (Brazilian Agricultural Research Corporation) developed the BR 473 maize variety, which has excellent energetic value and higher proteic content than the normal maize. BR 473 maize variety has 0.9 and 4.0 g/kg of grain of tryptophan and lysine, respectively, against the 0.6 and 2.6 from normal maize. The high lysine content in some maize varieties as opaque-2 (o2), for example, has been related to the presence of non-zein proteins. In order to identify them, which could explain the high proteic content of BR 473 maize variety, the protein bodies proteome of these grains were analyzed. For this purpose, one and two-dimensional liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS and LC/LC-MS/MS) were used and evaluated, since these techniques have become a powerful tool for protein sequencing and identification. We have identified six proteins by LC-MS/MS and sixteen proteins by LC/LC-MS/MS

First phytochemical study and biological activity of the leaves ethanolic extract from Cissus spinosa Cambess

AIMS: The objective of this study was to identify the phytochemical profile and to evaluate the biological effects of the crude ethanolic extract (EE) and the ethanolic fraction (EF) of leaves of the species Cissus spinosa Cambess, after oxidative stress induced by cyclophosphamide (CP) in mice.METHODS: Phytochemical profile was performed detecting functional groups and, analysis of total flavonoids and phenols concentration, as well as the antiradical activity in EE and EF. The phytochemical characterization was done for the identification of flavonoids present in the leaves of the plant. In the biochemical tests, hematological parameters, glucose and total cholesterol dosages in plasma, enzymatic and non-enzymatic antioxidants and lipid damage marker were evaluated in different tissues (liver, kidney and heart), besides genotoxic and immunological analyzes. The animals received 15 days of treatment, via gavage, with EE (50 mg kg-1) or EF (50 mg kg-1) and on the 15th day, an intraperitoneal injection of CP (100 mg kg-1) or saline (0.9%). After 24 h the last treatment, the animals were anesthetized for blood withdrawal, sacrificed and removal of the organs.RESULTS: In the phytochemical analyzes, the presence of alkaloids, flavonoids and phenols was identified, the latter presented a higher concentration for EF. Eight flavonoids were identified - Rutin, Quercetin-3-β-D-glucoside, Quercitrin, Taxifolin, Quercetin, Canferol, Luteolin and Apigenin. In the biochemical analyzes, in general, EE showed a better antioxidant action against oxidative damages, hypoglycemic and antitilipemic action when comparing with EF, probably due to the synergism caused by flavonoids. It was observed the reduction and an increase of micronucleated polychromatic erythrocytes, due to the action of antioxidant compounds and alkaloids present in the plant, also considering the question of the seasonal period that directly interferes in the production of these compounds. In the immunological analysis, the extracts did not stimulate the spontaneous production of oxygen peroxide (H2O2) and nitric oxide (NO•). CONCLUSIONS: Other studies, such as the variation of the chemical composition of the plant by local seasonality, hypoglycemic and antilipemic action, should be carried out to better delineate the biological action present in this plant.Objetivos: O objetivo deste estudo foi identificar o perfil fitoquímico e avaliar os efeitos biológicos do extrato etanólico bruto (EE) e da fração etanólica (EF) das folhas da espécie Cissus spinosa Cambess, após estresse oxidativo induzido pela ciclofosfamida. (CP) em camundongos.Métodos: O perfil fitoquímico foi realizado detectando grupos funcionais e, análise da concentração de flavo-noides e fenóis totais, bem como a atividade antirradical em EE e EF. A caracterização fitoquímica foi realizada para a identificação de flavonoides presentes nas fo-lhas da planta. Nos testes bioquímicos, os parâmetros hematológicos, as dosagens de glicose, colesterol total, creatinina, fosfatase alcalina e aminotransferases no plasma, os antioxidantes enzimáticos e não enzimá-ticos e o marcador de dano lipídico foram avaliados em diferentes tecidos (fígado, rim e coração), além de análises genotóxicas e imunológicas. Os animais receberam 15 dias de tratamento, via gavagem, com EE (50 mg kg-1) ou EF (50 mg kg-1) e no 15º dia, injeção intraperitoneal de CP (100 mg kg-1) ou soro fisiológico (0,9%) . Após 24 h do último tratamento, os animais fo-ram anestesiados para retirada do sangue, sacrificados e após a retirada dos órgãos.Resultados: Nas análises fitoquímicas, identificou-se a presença de alcaloides, flavonoides e fenóis, estes últimos apresentaram maior concentração para EF. Oito flavonoides foram identificados - rutina, quercetina-3-β--D-glicosídeo, quercitrina, taxifolina, quercetina, can-ferol, luteolina e apigenina. Nas análises bioquímicas, em geral, o EE apresentou melhor ação antioxidante contra os danos oxidativos, ação hipoglicemiante e antitilipêmica quando comparada à EF, provavelmente devido ao sinergismo causado pelos flavonoides. Observou-se a redução e um aumento de eritrócitos policromáticos micronucleados, devido à ação de compostos antioxidantes e alcaloides presentes na planta, considerando também a questão do período sazonal que interfere diretamente na produção desses compostos. Na análise imunológica, os extratos não estimularam a produção espontânea de peróxido de oxigênio (H2O2) e óxido nítrico (NO•).Conclusões: Outros estudos, como a variação da composição química da planta por sazonalidade local, ação hipoglicemiante e antilipêmica, devem ser rea-lizados para melhor delineamento da ação biológica presente nesta planta

Improved antimutagenic effect of Pyrostegia venusta (Ker Gawl.) Miers nanostructured extract in liposome and polymeric nanoparticle

Pyrostegia venusta (Ker Gawl.) Miers, popularly known as “Cipó-de São-João”, has been used in traditional medicine for its therapeutic properties. Nanotechnology is able to enhance the pharmacological activity of plant extracts. In this context, liposomes and polymeric nanoparticles containing P. venusta ethanolic extract were developed and then physico-chemically characterized to evaluate the mutagenic/antimutagenic effects of P. venusta. In addition, transaminases and serum creatinine were biochemically analyzed for liver and renal damage, respectively. The micronucleus test was performed with male Swiss mice treated orally for 15 consecutive days with free extracts and nanostructured with P. venusta, and then intraperitoneally with N-ethyl-N-nitrosurea (50 mg/kg) on the 15th day of treatment. Micronucleated polychromatic erythrocytes (MNPCE) were evaluated in bone marrow. There was a significant reduction in the frequency of MNPCE (LPEPV = 183% and NPEPV = 114%, p < 0.001), indicating antimutagenic potential of the nanostructured extracts with P. venusta. The groups treated with only nanostructured extract did not show an increase in MNPCE frequency, and biochemical analyzes showed no significant difference between treatments. The liposomes and polymeric nanoparticles containing Pyrostegia venusta ethanolic extract showed biological potential in preventing the first step of carcinogenesis under the experimental conditions

Antioxidant potential of Carica papaya Linn (Caricaceae) leaf extract in mice with cyclophosphamide induced oxidative stress

AIMS: This study aimed to investigate the effects of crude extract of Carica papaya leaves on oxidative stress in mice induced by cyclophosphamide, as well as phytochemical profile characterization of this extract.METHODS: The male Swiss mice received 15 days of treatment with the extract (500 mg kg-1, via gavage) and intraperitoneal injection of cyclophosphamide (75 mg kg-1) or saline (0.9%) on the 15th day. After 24 h the last treatment, the animals were anesthetized for blood withdrawal, sacrificed and removal of the organs for analyses (liver, kidney and heart). In the biochemical tests were determined: hematological parameters in blood, aminotransferases, alkaline phosphatase, glucose and total cholesterol dosages in plasma, enzymatic and non-enzymatic antioxidants and lipid damage marker were evaluated in different tissues, besides genotoxic and histopathological analyzes.RESULTS: In the extract of Carica papaya leaves, the flavonoids quercetin-3β-D-glucoside and rutin were identified, besides present positive results for alkaloids, saponins and tannins. This extract increased the activity of glutathione-S-transferase and catalase enzymes in the liver and reduced the levels of reduced glutathione in the kidneys and hematocrit levels, red cell count, and hemoglobin. It promoted the decrease of the reactive species of thiobarbituric acid (TBARS) in the kidneys and the activity of enzyme aspartate aminotransferase in the plasma and was antimutagenic in the micronucleus test.CONCLUSIONS: The study showed that extract of Carica papaya was beneficial against oxidative events and prevented DNA damage. The extract also showed hepatotoxicity, therefore prolonged infusion of papaya leaves is not advisable.OBJETIVOS: O objetivo deste estudo foi investigar os efeitos do extrato bruto de folhas de Carica papaya sobre o estresse oxidativo em camundongos induzidos pela ciclofosfamida, bem como a caracterização do perfil fitoquímico deste extrato.MÉTODOS: Os camundongos Swiss machos receberam 15 dias de tratamento com o extrato (500 mg kg-1, via gavagem) e injeção intraperitoneal de ciclofosfamida (75 mg kg-1) ou salina (0,9%) no 15º dia. Após 24 h do último tratamento, os animais foram anestesiados para retirada do sangue, sacrificados e retirada dos órgãos para análises (fígado, rim e coração). Nos testes bioquímicos foram determinados: parâmetros hematológicos em sangue, aminotransferases, fosfatase alcalina, dosagens de glicose e colesterol total no plasma, antioxidantes enzimáticos e não enzimáticos e marcador de dano lipídico foram avaliados em diferentes tecidos, além de análises genotóxicas e histopatológicas.RESULTADOS: No extrato de folhas de Carica papaya foram identificados os flavonoides quercetina-3β-D-glicosídeo e rutina, além de resultados positivos para alcaloides, saponinas e taninos. Este extrato aumentou a atividade das enzimas glutationa-S-transferase e catalase no fígado e diminuiu os níveis de glutationa reduzida nos rins, a concentração do hematócrito, a contagem dos glóbulos vermelhos e a hemoglobina. Promoveu a diminuição das espécies reativas de ácido tiobarbitúrico (TBARS) nos rins, a atividade da enzimaaspartato aminotransferase no plasma e foi antimutagênico no teste do micronúcleo.CONCLUSÕES: O estudo mostrou que o extrato de Carica papaya foi benéfico contra eventos oxidativos e preveniu danos no DNA. O extrato também mostrou hepatotoxicidade, portanto, a infusão prolongada de folhas de mamão não é aconselhável

Coronary revascularization with the left internal thoracic artery and radial artery: comparison of short-term clinical evolution between elective and emergency surgery

BACKGROUND: Left internal thoracic artery-to left anterior descending artery grafting has become a fundamental part of coronary artery bypass grafting. This grafting has led to increased use of other arterial conduits, of which the radial artery is most popular. Whether radial grafting can be used in the emergency patient is not known. This study compares the short-term clinical evolution between elective vs emergency coronary artery bypass grafting surgery with left internal thoracic artery and radial artery. METHODS: A retrospective study of 47 patients who underwent elective or emergency coronary artery bypass grafting from 1996 to 2003. All patients had coronary stenosis >70% in all target vessels. Only the left internal thoracic artery and radial artery were used as grafts. Patients were divided into elective group (23 patients) and emergency group (24 patients). Emergency criteria were unstable angina and/or critical coronary stenosis with high risk for acute myocardial infarction. Groups were similar for age and number of diseased vessels. RESULTS: The mean number of left internal thoracic artery grafts per patient in the elective and emergency groups were respectively 1.17 and 1.38 (P = .17). The mean number of radial artery grafts per patient in the elective and emergency groups was respectively 2.26 and 2.08 (P = .48). The 30-day mortality was 0. There was no postoperative cardiogenic shock. The elective group had 1 acute myocardial infarction (4.4%) postoperatively, and emergency group had 5 (20.8%). A nonsignificant trend towards acute myocardial infarction was noted in the emergency group (P = .18). Intensive care unit and postoperative stay were similar in both groups. CONCLUSION: Coronary artery bypass grafting using left internal thoracic artery and radial artery accomplishing complete revascularization can be performed in emergency patients with results similar to those for elective patients

Brazilian Flora 2020: Leveraging the power of a collaborative scientific network

International audienceThe shortage of reliable primary taxonomic data limits the description of biological taxa and the understanding of biodiversity patterns and processes, complicating biogeographical, ecological, and evolutionary studies. This deficit creates a significant taxonomic impediment to biodiversity research and conservation planning. The taxonomic impediment and the biodiversity crisis are widely recognized, highlighting the urgent need for reliable taxonomic data. Over the past decade, numerous countries worldwide have devoted considerable effort to Target 1 of the Global Strategy for Plant Conservation (GSPC), which called for the preparation of a working list of all known plant species by 2010 and an online world Flora by 2020. Brazil is a megadiverse country, home to more of the world's known plant species than any other country. Despite that, Flora Brasiliensis, concluded in 1906, was the last comprehensive treatment of the Brazilian flora. The lack of accurate estimates of the number of species of algae, fungi, and plants occurring in Brazil contributes to the prevailing taxonomic impediment and delays progress towards the GSPC targets. Over the past 12 years, a legion of taxonomists motivated to meet Target 1 of the GSPC, worked together to gather and integrate knowledge on the algal, plant, and fungal diversity of Brazil. Overall, a team of about 980 taxonomists joined efforts in a highly collaborative project that used cybertaxonomy to prepare an updated Flora of Brazil, showing the power of scientific collaboration to reach ambitious goals. This paper presents an overview of the Brazilian Flora 2020 and provides taxonomic and spatial updates on the algae, fungi, and plants found in one of the world's most biodiverse countries. We further identify collection gaps and summarize future goals that extend beyond 2020. Our results show that Brazil is home to 46,975 native species of algae, fungi, and plants, of which 19,669 are endemic to the country. The data compiled to date suggests that the Atlantic Rainforest might be the most diverse Brazilian domain for all plant groups except gymnosperms, which are most diverse in the Amazon. However, scientific knowledge of Brazilian diversity is still unequally distributed, with the Atlantic Rainforest and the Cerrado being the most intensively sampled and studied biomes in the country. In times of “scientific reductionism”, with botanical and mycological sciences suffering pervasive depreciation in recent decades, the first online Flora of Brazil 2020 significantly enhanced the quality and quantity of taxonomic data available for algae, fungi, and plants from Brazil. This project also made all the information freely available online, providing a firm foundation for future research and for the management, conservation, and sustainable use of the Brazilian funga and flora